Abstract
The effects of biofilm growth and methane gas generation on water flow in porous media were investigated in an anaerobic two-dimensional sand-filled cell. Inoculation of the lower portion of the cell with a methanogenic culture and addition of methanol to the bottom of the cell led to biomass growth and formation of a gas phase. Biomass distributions in the water and on the sand in the cell were measured by protein analysis. The biofilm distribution on sand was observed by confocal laser scanning microscopy. The formation, migration, distribution and saturation of gases in the cell were visualized by the charge-coupled device (CCD) camera. The effects of biofilm and gas generation on water flow were separated by performing one tracer test in the presence of both biofilm and a gas phase and a second tracer test after removal of the gas phase through water flushing. The results of tracer tests demonstrated that flow and transport in the two-dimensional cell were significantly affected by both gas generation and biofilm growth. Gas generated at the bottom of the cell in the biologically active zone moved upwards in discrete fingers, so that gas phase saturations (gas-filled fraction of void space) in the biologically active zone at the bottom of the cell did not exceed 40–50%, while gas accumulation at the top of the cell produced gas phase saturations as high as 80%. The greatest reductions in water phase permeability, based on measurements of reductions in water phase saturations, occurred near the top of the box as a result of the gas accumulation. In contrast the greatest reductions in permeability due to biofilm growth, based on measurements of biofilm thickness, occurred in the most biologically active zone at the bottom of the cell, where gas phase saturations were approximately 40–50%, but permeability reductions due to biofilm growth were estimated to be 80–95%.
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